• Title/Summary/Keyword: Indicator assay

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Correlation between Cerebral Vasospasm after Subarachnoid Hemorrhage and Intercellular Adhesion Molecule-1 Levels in Serum and Cerebrospinal Fluid

  • Cheong, Jin-Hwan;Kim, Jae-Min;Bak, Koang-Hum;Kim, Choong-Hyun
    • Journal of Korean Neurosurgical Society
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    • v.38 no.1
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    • pp.1-11
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    • 2005
  • Objective : The purpose of this study is to evaluate the correlation of the ICAM-l levels in serum and CSF with cerebral vasospasm in early aneurysmal subarachnoid hemorrhage [SAH] patients. Methods : A prospective analysis was performed in thirty consecutive patients who underwent early surgery for intracranial aneurysmal SAH. The serum and CSF were obtained daily through the indwelling arterial lines and intraoperative ventriculostomy, or cisternal drain for 4 consecutive days after surgery. The ICAM-1 levels in serum and CSF samples were measured via quantitative enzyme-linked immunosorbent assay. Results : The mean concentration of serum in aneurysmal SAH patients was 207.89ng/ml compared with 132.25ng/ml in controls. The mean concentration of CSF in aneurysmal SAH patients was 76.39ng/ml compared with 3.96ng/ml in controls. There were no significant differences between serum and CSF ICAM-1 level with regards to clinical characteristics in patients with aneurysmal SAH [P>0.05]. However, CSF ICAM-1 levels increased significantly in patients with vasospasm compared with those without vasospasm [P<0.05]. Conclusion : The major result of this study shows that ICAM-1 is increased in CSF after early aneurysmal SAH and that this increase in ICAM-1 has correlation with cerebral vasospasm. Further study is needed to determine whether ICAM-1 levels may be indicator in the pathogenesis of important events leading to cerebral vasospasm.

Arginase inhibition by rhaponticin increases L-arginine concentration that contributes to Ca2+-dependent eNOS activation

  • Koo, Bon-Hyeock;Lee, Jonghoon;Jin, Younghyun;Lim, Hyun Kyo;Ryoo, Sungwoo
    • BMB Reports
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    • v.54 no.10
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    • pp.516-521
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    • 2021
  • Although arginase primarily participates in the last reaction of the urea cycle, we have previously demonstrated that arginase II is an important cytosolic calcium regulator through spermine production in a p32-dependent manner. Here, we demonstrated that rhaponticin (RPT) is a novel medicinal-plant arginase inhibitor and investigated its mechanism of action on Ca2+-dependent endothelial nitric oxide synthase (eNOS) activation. RPT was uncompetitively inhibited for both arginases I and II prepared from mouse liver and kidney. It also inhibited arginase activity in both aorta and human umbilical vein endothelial cells (HUVECs). Using both microscope and FACS analyses, RPT treatments induced increases in cytosolic Ca2+ levels using Fluo-4 AM as a calcium indicator. Increased cytosolic Ca2+ elicited the phosphorylations of both CaMKII and eNOS Ser1177 in a time-dependent manner. RPT incubations also increased intracellular L-arginine (L-Arg) levels and activated the CaMKII/AMPK/Akt/eNOS signaling cascade in HUVECs. Treatment of L-Arg and ABH, arginase inhibitor, increased intracellular Ca2+ concentrations and activated CaMKII-dependent eNOS activation in ECs of WT mice, but, the effects were not observed in ECs of inositol triphosphate receptor type 1 knockout (IP3R1-/-) mice. In the aortic endothelium of WT mice, RPT also augmented nitric oxide (NO) production and attenuated reactive oxygen species (ROS) generation. In a vascular tension assay using RPT-treated aortic tissue, cumulative vasorelaxant responses to acetylcholine (Ach) were enhanced, and phenylephrine (PE)-dependent vasoconstrictive responses were retarded, although sodium nitroprusside and KCl responses were not different. In this study, we present a novel mechanism for RPT, as an arginase inhibitor, to increase cytosolic Ca2+ concentration in a L-Arg-dependent manner and enhance endothelial function through eNOS activation.

Serum Concentration of Nitrotyrosine as Indicator of Disease Progress in Dogs with Myxomatous Mitral Valve Disease

  • Bang, Ju-Hwan;Park, Jun-Seok;Seo, Kyoung-Won;Song, Kun-Ho
    • Journal of Veterinary Clinics
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    • v.36 no.2
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    • pp.102-105
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    • 2019
  • Nitrotyrosine was found to be dependent on the severity of myxomatous mitral valve disease (MMVD). However, a correlation of serum nitrotyrosine concentration in dogs with MMVD and the progression of the disease has not been investigated. This study compared changes in serum nitrotyrosine concentration with the progression of MMVD. Nine client-owned dogs were recruited for the study. Dogs were classified by measuring the amount of regurgitation using echocardiography into mild, moderate, or severe MMVD groups. Serum nitrotyrosine concentration was measured by an enzyme-linked immunosorbent assay test. Serum nitrotyrosine concentration was significantly higher at 180 days than at 0 day (P < 0.05). However, serum nitrotyrosine concentration at 360 days was lower than that at 180 days (P < 0.05). Serum nitrotyrosine concentration at 540 days was lower than at 180 days (P < 0.05). There was no correlation between serum nitrotyrosine and left atrial to aortic root diameter ratio (LA/Ao ratio) (n = 33, $R^2=0.003$, P = 0.759). Also, there was no correlation between serum nitrotyrosine and vertebral heart score (VHS) (n = 33, $R^2=0.026$, P = 0.368) and left ventricular end-diastolic diameter, normalized for body weight by the formula (LVEDDN) (n = 33, $R^2=0.053$, P = 0.196). The results of the study suggest that the progression of MMVD is correlated with changes in serum nitrotyrosine concentration, which shows potential for use as a cardiac biomarker which can be used to analyze the progression of disease in MMVD.

KAT8/MOF-Mediated Anti-Cancer Mechanism of Gemcitabine in Human Bladder Cancer Cells

  • Zhu, Huihui;Wang, Yong;Wei, Tao;Zhao, Xiaoming;Li, Fuqiang;Li, Yana;Wang, Fei;Cai, Yong;Jin, Jingji
    • Biomolecules & Therapeutics
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    • v.29 no.2
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    • pp.184-194
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    • 2021
  • Histone acetylation is a well-characterized epigenetic modification controlled by histone acetyltransferases (HATs) and histone deacetylases (HDACs). Imbalanced histone acetylation has been observed in many primary cancers. Therefore, efforts have been made to find drugs or small molecules such as HDAC inhibitors that can revert acetylation levels to normal in cancer cells. We observed dose-dependent reduction in the endogenous and exogenous protein expression levels of KAT8 (also known as human MOF), a member of the MYST family of HATs, and its corresponding histone acetylation at H4K5, H4K8, and H4K16 in chemotherapy drug gemcitabine (GEM)-exposed T24 bladder cancer (BLCA) cells. Interestingly, the reduction in MOF and histone H4 acetylation was inversely proportional to GEM-induced γH2AX, an indicator of chemotherapy drug effectiveness. Furthermore, pGL4-MOF-Luc reporter activities were significantly inhibited by GEM, thereby suggesting that GEM utilizes an MOF-mediated anti-BLCA mechanism of action. In the CCK-8, wound healing assays and Transwell® experiments, the additive effects on cell proliferation and migration were observed in the presence of exogenous MOF and GEM. In addition, the promoted cell sensitivity to GEM by exogenous MOF in BLCA cells was confirmed using an Annexin V-FITC/PI assay. Taken together, our results provide the theoretical basis for elucidating the anti-BLCA mechanism of GEM.

Investigation of Pharmacological Stability of Hyeonggaeyeongyotang Gagambang by Formulation Changes for Foaming Tablet (형개연교탕(荊芥連翹湯) 가감방(가감방)의 발포정 제형 변화에 따른 약리학적 안정성 연구)

  • Cho, Nam Joon;Gang, Seong Gu;Kim, Ji Young;Han, Hyo Sang;Kim, Kee Kwang
    • The Korea Journal of Herbology
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    • v.33 no.5
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    • pp.67-72
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    • 2018
  • Objectives : Natural extracts have been extensively studied to replace single agent drugs that cause a variety of side effects. However, studies of changes to the formulation of natural extracts has not been nearly proceed. We aimed to investigate whether pharmacological stability of hyeonggaeyeongyotang gagambang (HYT) is altered by formulation changes for foaming tablet. Methods : In this study, we performed freeze - drying of HYT, which is known to have antioxidant and anti - inflammatory properties, and then changed the formulation by foaming. Results : As a result, the foaming reaction appeared normally when HYT foamed tablets were put into water, and almost all of the substances were dissolved in the aqueous solution. In addition, we confirmed using high-performance liquid chromatograph that the geniposide used as an indicator material of HYT was stable in most of the formulations. It was confirmed that the change of HYT formulation did not affect the antioxidant efficacy by the 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid assay. Moreover, quantitative real-time PCR confirmed that the inhibitory effect of HYT on IL-$1{\beta}$ mRNA expression induced by lipopolysaccharides treatment in murine macrophage RAW 264.7 cells was similar in the solution of foaming tablet. Conclusions : These results suggest that the materials with various pharmacological effects can be stably maintained even when the formulation is changed by the foaming action of HYT. Our results are expected to provide important basic knowledge on formulation changes using various natural extracts.

Biological and Molecular Characterization of a Korean Isolate of Orthotospovirus chrysanthinecrocaulis (Formerly Chrysanthemum Stem Necrosis Virus) Isolated from Chrysanthemum morifolium

  • Seong Hyeon Yoon;Su Bin Lee;Eseul Baek;Ho-Jong Ju;Ju-Yeon Yoon
    • Research in Plant Disease
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    • v.29 no.3
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    • pp.286-294
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    • 2023
  • Biological and molecular characterization of a Korean isolate of Orthotospovirus chrysanthinecrocaulis (formerly known as chrysanthemum stem necrosis virus, CSNV) isolated from Chrysanthemum morifolium was determined using host range and sequence analysis in this study. Twenty-three species of indicator plants inoculated mechanically CSNV-Kr was investigated for determination of host range. CSNV-Kr induced various local and systemic symptoms in the inoculated plant species. CSNV-Kr could not infect three plant species and induced symptomless in systemic leaves in Nicotiana tabacum cultivars, though the plant samples reacted positively with the antiserum to CSNV by double-antibody sandwich-enzyme-linked immunosorbent assay. The complete genome sequence of CSNV-Kr was determined. The L RNA of CSNV-Kr consists of 8,959 nucleotides (nt) and encodes a putative RNA-dependent RNA polymerase. The M RNA of CSNV-Kr consists of 4,835 nt and encodes the movement protein (NSm) and the glycoprotein precursor (Gn/Gc protein). The S RNA of CNSV-Kr consists of 2,836 nt and encodes NSs protein and N protein. The Gn/Gc and N sequence of CSNV-Kr were compared with those of previously published CSNV isolates originating from different countries at nucleotide and amino acid levels. The Gn/GC sequence of CSNV-Kr shared 98.8-99.5% identity with CSNV isolated from other countries and the N sequence of CSNV-Kr shared 98.8-99.6% identity. No particular region of variability could be found in either grouping of viruses. All of the CSNV isolates did not show any relationship according to geographical origins and isolation hosts, suggesting no distinct segregation of the CSNV isolates.

A Clinical Study of InGaAlP Laser Transcutaneous Blood Irradiation on Heart Rate Variability in Healthy Adults (II) (InGaAlP 레이저 경피혈액조사가 정상성인의 심박변이도에 미치는 영향에 대한 임상적 연구(II))

  • Yeo, Jinju;Lee, Taeho;Son, Donghyuk;Hsing, Lichang;Lee, Inhwan;Jang, Insoo
    • The Journal of the Society of Stroke on Korean Medicine
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    • v.6 no.1
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    • pp.9-16
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    • 2005
  • Objective : The heart rate variability is very useful indicator to study the function of the autonomic nervous system(ANS), and the physiologic signals can be observed based on the changes of the ANS of the heart. In order to assay the effects of the laser exposing to healthy subjects, the double blind test has been performed. Methods : This study included 62 healthy adults who have not any ANS disease and had normal sinus rhythm in electrocardiogram. The control group consisted of 31 subjects, laser group consisted of 31 subject. HRV was measured for 5 minutes before laser irradiation, sham and real laser irradiated for 30 minutes and than HRV remeasured for 5 minutes. Statistical significance was evaluated by independent T-test. Results : Mean HRV, Ln(VLF), Ln(HF), Ln(TP) of both groups at post-laser period decreased compared with that of the pre-laser period. Ln(LF) of both groups at post-laser period increased compared with that of the pre-laser period. LF/HF, SDNN of real laser group decreased and sham group decreased. Conclusions : There is no difference between two groups. The reason is presumed that all the studied subjects are healthy adults, and also the short and single transcutaneous laser irradiation would not influence upon changes of the ANS. The further study must be followed.

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Validation of an Analytical Method for Deacetylasperulosidic acid, Total Sugar and Monosaccharide Analysis in Fermented Morinda citrifolia Polysaccharide Powder (발효노니 다당체 분말의 deacetylasperulosidic acid, 총당 및 단당류 분석법 검증)

  • Kwon, Heeyeon;Choi, Jisoo;Kim, Soojin;Kim, Eunmin;Uhm, Jihyun;Kim, Bokyung;Lee, Jaeyeon;Kim, Yongdeok
    • Journal of Food Hygiene and Safety
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    • v.37 no.4
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    • pp.216-224
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    • 2022
  • This study was aimed at validating the analysis methods for deacetylasperulosidic acid (DAA), total sugar, galacturonic acid, glucose, and galactose, which are the indicator components of fermented Morinda citrifolia polysaccharide extract (Vitalbos). We modified the previously reported methods for validating the analytical methods. The specificity, linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) were measured using phenol-sulfuric acid method and high-performance liquid chromatography (HPLC). The retention time and spectrum of the standard solution of Vitalbos coincided, confirming the specificity. The calibration curve correlation coefficient (R2), of five indicator components, ranged from 0.9995-0.9998, indicating excellent linearity of 0.99 or more. The intra-day and inter-day precision range of the assay was 0.14-3.01%, indicating a precision of less than 5%. The recovery rate was in the range of 95.13-105.59%, presenting excellent accuracy. The LOD ranged from 0.39 to 0.84 ㎍/mL and the LOQ ranged from 1.18 to 2.55 ㎍/mL. Therefore, the analytical method was validated for DAA, total sugar, galacturonic acid, glucose, and galactose, in Vitalbos. The indicator component content in Vitalbos was determined using a validated method. The contents of DAA, total sugar, galacturonic acid, glucose, and galactose were 2.31±0.06, 475.92±5.95, 72.83±1.05, 71.63±2.44, and 67.30±2.31 mg/g of dry weight, respectively. These results suggest that the developed analytical method is efficient and could contribute to the quality control of Vitalbos, as a healthy functional food material.

Correlation between Antioxidant Capacities and Color Values in Korean Red Grape Juices (국내 적포도주스의 항산화능과 색도의 상관성)

  • Kim, Joo-Young;Seong, Gi-Un;Hwang, In-Wook;Chung, Shin-Kyo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.8
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    • pp.1206-1211
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    • 2015
  • Grape juice is consumed worldwide and studied due to the high antioxidant activities and contents. The color of grape juice is due to the presence of phenolic compounds such as anthocyanins. Therefore, color values can be an indicator of antioxidant capacities of grape juice. However, the correlation between color values and antioxidant activities in grape juice has not been well studied. In this study, the physicochemical characteristics and correlation between color values and antioxidant capacities of Korean red grape juices (five commercial juices from market and two juices prepared in the laboratory) were investigated to estimate antioxidant capacities. Antioxidant capacities were determined by 1,1-diphenlyl-2-picrylhydrazyl radical scavenging assay, ferric ion reducing antioxidant power assay, and oxygen radical absorbance capacity assay. Total phenolic contents, total flavonoid contents, and total anthocyanin contents, including five kinds of polyphenolic compounds, were examined by high performance liquid chromatography (HPLC). The results for physicochemical properties showed similar values, except titratable acidity. The color hue values of the prepared juices were higher than those of commercial juices, which was in contrast to the lower color intensity values (P<0.05). The Hunter L, a, and b values showed almost no difference between commercial and prepared juices. The antioxidant activities and total phenolic contents of commercial juices were higher than those of prepared ones. Gallic acid, catechin, and quercetin were confirmed by HPLC in all samples. The total phenolic and total flavonoid contents showed positive correlation with antioxidant activities. In addition, antioxidant activities and contents correlated with color values. Thus, estimation of antioxidant capacity could be feasible through the spectrophotometric measurement of color values.

Induction of Apoptosis in Human Cancer Cells with Extracts of Taraxacum coreanum, Youngia sonchifolia and Ixeris dentate (흰민들레, 고들빼기, 씀바귀 추출물의 암세포 증식 억제 효과)

  • Shin, Seong-Ah;Lee, Hae-Nim;Choo, Gang-Sik;Kim, Hyeong-Jin;Park, Byung-Kwon;Kim, Byeong-Soo;Jung, Ji-Youn
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.51-58
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    • 2016
  • This research examined the effects of Compositae extract on the inhibition of proliferation and apoptosis in human breast and human gastric cancer cells. Compositae extracts which is used in the experiment are Taraxacum coreanum (TC), Youngia sonchifolia (YS) and Ixeris dentata (ID). The proliferation of SK-BR-3, MDA-MB-231 and AGS cells were investigated by MTT assay. ID and YS extracts inhibited proliferation of SK-BR-3, MDA-MB-231 and AGS cells in a dose-dependent manner, but TC have barely affected. In addition, the most effective extract was ID. To assess the apoptosis of ID extract, the nuclei of human cancer cells were stained with DAPI solution respectively. Chromatin condensation, indicated apoptosis, was increased in a dose-dependent manner. We investigated change of ID extract-induced apoptosis proteins on human cancer cells by western blot analysis. The level of Bcl-2 decreased, whereas the level of Bax, cleaved-PARP increased in dose-dependent manner compared with non-treatment. Also Bax/Bcl-2 ratio, which is used in clinical indicator of apoptosis, was increased at ID extract treatment group compared with non-treatment. Moreover the Bax/Bcl-2 ratio of MDA-MB-231 cell was significantly increased as against SK-BR-3, AGS cells. These results indicated that ID extract have anti-proliferation effect better than YS or TC, and induced apoptosis in human breast cancer MDA-MB-231 cell better than human breast cancer SK-BR-3 cell, human gastric cancer. Even if further research is needed, ID can be developed as a chemopreventive or therapeutic agent of breast cancer.